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. 2023 Jan 29;15(2):113.
doi: 10.3390/toxins15020113.

Aggregation-Induced Red Emission Nanoparticle-Based Lateral Flow Immunoassay for Highly Sensitive Detection of Staphylococcal Enterotoxin A

Hanpeng Xiong  1   2 , Ping Chen  1   2 , Xirui Chen  1   2 , Xuanang Shen  1   2 , Xiaolin Huang  1   2   3 , Yonghua Xiong  1   2   3 , Yu Su  1   3
Affiliations

Aggregation-Induced Red Emission Nanoparticle-Based Lateral Flow Immunoassay for Highly Sensitive Detection of Staphylococcal Enterotoxin A

Hanpeng Xiong et al. Toxins (Basel). .

Abstract

Staphylococcal enterotoxin A (SEA) has presented enormous difficulties in dairy food safety and the sensitive detection of SEA provides opportunities for effective food safety controls and staphylococcal food poisoning tracebacks. Herein, a novel aggregation-induced emission (AIE)-based sandwich lateral flow immunoassay (LFIA) was introduced to detect SEA by using red-emissive AIE nanoparticles (AIENPs) as the fluorescent nanoprobe. The nanoprobe was constructed by directly immobilising antibodies on boronate-tagged AIENPs (PBA-AIENPs) via a boronate affinity reaction, which exhibited a high SEA-specific affinity and remarkable fluorescent performance. Under optimal conditions, the ultrasensitive detection of SEA in pasteurised milk was achieved within 20 min with a limit of detection of 0.04 ng mL-1. The average recoveries of the PBA-AIENP-LFIA ranged from 91.3% to 117.6% and the coefficient of variation was below 15%. It was also demonstrated that the PBA-AIENP-LFIA had an excellent selectivity against other SE serotypes. Taking advantage of the excellent sensitivity of this approach, real chicken and salad samples were further analysed, with a high versatility and accuracy. The proposed PBA-AIENP-LFIA platform shows promise as a potent tool for the identification of additional compounds in food samples as well as an ideal test method for on-site detections.

Keywords: aggregation-induced emission; boronate affinity reaction; lateral flow immunoassay; staphylococcal enterotoxin A.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Figures

Scheme 1
Scheme 1
Schematic illustration of (A) fabrication of AIENPs@mAbs and (B) construction of sandwich PBA-AIENP-LFIA platform for SEA detection.
Figure 1
Figure 1
(A) TEM and (B) SEM images of AIENPs. (C) UV-vis absorption spectrum and fluorescence emission spectrum of AIENPs. (D) Zeta potential evolution and (E) fluorescence emission spectra of AIENPs and PBA-AIENPs. (F) Dynamic light scattering analysis of AIENPs and PBA-AIENPs@mAbs.
Figure 2
Figure 2
(A) Effect of pH on the coupling of anti-SEA mAbs and PBA-AIENPs. The added mAbs concentration was 20 µg mL−1. (B) The added amount of labelled anti-SEA mAbs on the coupling of anti-SEA mAbs and PBA-AIENPs at a pH of 7.0. (C) Effect of the added amount of labelled anti-SEA mAbs on the conjugation of anti-SEA mAbs and AIENPs by EDC method. The spiked SEA concentration was 20 ng mL−1.
Figure 3
Figure 3
Quantitative detection of SEA via the proposed PBA-AIENP-LFIA platform. (A) Stereograms of PBA-AIENP-LFIA for SEA-spiked milk samples with SEA concentrations of 0, 0.02, 0.05, 0.1, 0.2, 0.5, 1, 2, 5, 10, 20, 40, 80, 100, 120 and 150 ng mL−1. (B) FIT/FIC signal of the PBA-AIENP-LFIA platform at SEA concentrations ranging from 0 to 500 ng mL−1 in pasteurised milk. (C) Standard curve of PBA-AIENP-LFIA for SEA determination in pasteurised milk samples.
Figure 4
Figure 4
Evolution of the specificity of the PBA-AIENP-LFIA to staphylococcal enterotoxins (SEA, SEB, SEC, SED and SEE).
See this image and copyright information in PMC

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